| 杨光勇,郭海涛,刘茜明,何光志,田维毅,蔡琨,王平,梁建东,王文佳,韩洁.重组人可溶性白细胞介素4受体蛋白抑制大鼠哮喘[J].浙江中西医结合杂志,2016,26(4): |
| 重组人可溶性白细胞介素4受体蛋白抑制大鼠哮喘 |
| Recombinant soluble human interleukin-4 receptor protein inhibiting rat asthma model |
| 投稿时间：2015-11-08 修订日期：2016-01-20 |
| DOI： |
| 中文关键词: 重组人可溶性白细胞介素4受体 重组蛋白 哮喘模型 大鼠 |
| 英文关键词:Recombinant soluble human interleukin-4 receptor recombinant proteins Asthma model rat |
| 基金项目:2011 年贵州省社会发展科技攻关项目[黔科合 SY 字( 2011) 3020]、2014年度贵阳中医学院“2011协同创新中心”和贵州省委组织部高层次人才科研条件特助经费项目( TZJF - 2011 年28 号) 联合资助 |
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| 中文摘要: |
| 【】 目的 探讨重组可溶性白介素-4受体对大鼠哮喘模型的影响。方法 根据GenBank 中公布的Homo sapiens(human) IL-4R基因序列(XM_005255309) ,委托上海吉尔吉斯生物工程有限公司合成srhIL-4R 基因片段,将合成基因片段插入pET-28a( )构建重组表达载体pET-28a( )-srhIL-4R,酶切鉴定后,将载体转入表达菌株BL21(DE3)进行诱导表达重组蛋白srhIL-4R,并用SDS-PAGE和Western-blotting方法分析重组蛋白。提取重组蛋白干预哮喘模型大鼠,取肺组织进行病理切片观察。结果 重组载体pET-28a( )-srhIL-4R酶切获得目的片段长度为500bp～750bp之间的片段,表达载体pET-28a( )-srhIL-4R转入表达菌BL21(DE3) 后经IPTG诱导表达出的重组蛋白经SDS-PAGE和Western-blotting方法可见重组蛋白分子量约为25kDa；重组人可溶性IL-4R蛋白干预哮喘大鼠可见炎细胞数量较模型组明显减少。结论 srhIL-4R防治哮喘的具有一定功效,值得临床推广。 |
| 英文摘要: |
| [Abstract] Objective To investigate the effect of recombinant soluble human interleukin-4 receptor (srhIL-4R) in the asthma model. Methods The Homo sapiens (human) interleukin-4 receptor gene was synthesized according to the sequence of the GenBank (XM_005255309) by the Shanghai kyrgyzstan biological engineering co., LTD., the synthesis of gene fragment was cloned into pET-28a( ) vector to construct the recombinant expression vector pET-28a( )-srhIL-4R. The expression plasmid was induced into E. coli BL21 (DE3) and expression strain was selected, the recombinant was analyzed by SDS-PAGE and Western-blotting. Rat using an ovalbumin (OVA) -induced model of allergic asthma were conducted with the recombinant soluble human interleukin-4 receptor proteins, and lung tissue from model of allergic asthma were taken for pathology observation. Results We got 500bp~750bp target fragment by restricted enzyme, SDS-PAGE, and western-blotting analysis of the recombinant protein was about 25kDa. Compared with model group, allergic asthma intervention decreased significantly in inflammatory cells. Conclusion SrhIL-4R has a certain effect in the prevention and treatment of asthma, is worthy of clinical promotion. |
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